LOCUS       sequence-224271-        8430 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression construct .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8430)
  AUTHORS   Matsushime H, Ewen ME, Strom DK, Kato JY, Hanks SK, Roussel MF, 
            Sherr CJ
  TITLE     Identification and properties of an atypical catalytic subunit 
            (p34PSK-J3/cdk4) for mammalian D type G1 cyclins.
  JOURNAL   Cell. 1992 Oct 16;71(2):323-34.
  PUBMED    1423597
REFERENCE   2  (bases 1 to 8430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell."; 
            date: "1992-10-16"; volume: "71(2)"; pages: "323-34"
COMMENT     Sequence Label: pSRalpha-MSV-mouse Cdk4wt-Tk-neo
FEATURES             Location/Qualifiers
     source          1..8430
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     LTR             392..567
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    630..829
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     promoter        3050..3112
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             3169..3963
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3223..3242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3833..3852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5288..5422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5312..5331)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5366..5385
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5565..5587)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5728..5747
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5862..5879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6033..6621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6113..6132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6792..7583,7584..7652))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7415..7434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7653..7757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7825..7843
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7917..7937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7922..8251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8102..8237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaacat aactgagaat
       61 agaaaagttc agatcaaggt caggaacaaa gaaacagctg aataccaaac aggatatctg
      121 tggtaagcgg ttcctgcccc ggctcagggc caagaacaga tgagacagct gagtgatggg
      181 ccaaacagga tatctgtggt aagcagttcc tgccccggct cggggccaag aacagatggt
      241 ccccagatgc ggtccagccc tcagcagttt ctagtgaatc atcagatgtt tccagggtgc
      301 cccaaggacc tgaaaatgac cctttacctt atttgaacta accaatcagt tcgcttctcg
      361 cttctgttcg cgcgcttccg ctctccgagc tcaataaaag agcccacaac ccctcactcg
      421 gcgcgccagt cttccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct
      481 tgctgtttgc atccgaatcg tggtctcgct gttccttggg agggtctcct ctgagtgatt
      541 gactacccac gacgggggtc tttcatttgg gggctcgtcc gggatttgga gacccctgcc
      601 cagggaccac cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc
      661 gattgtctag tgtctatgtt tgatgttatg cgcctgcgtc tgtactagtt agctaactag
      721 ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct
      781 gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg
      841 atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag
      901 ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg
      961 cggaattcgg cacgagccct gcttcgagaa tggctgccac tcgatatgaa cccgtggctg
     1021 aaattggtgt cggtgcctat gggacggtgt acaaagcccg agatccccac agtggccact
     1081 ttgtggccct caagagtgtg agagttccta atggaggagc agctggaggg ggccttcccg
     1141 tcagcacagt tcgtgaggtg gccttgttaa ggaggctgga ggcctttgaa catcccaatg
     1201 ttgtacggct gatggatgtc tgtgctactt cccgaactga tcgggacatc aaggtcaccc
     1261 tagtgtttga gcatatagac caggacctga ggacatacct ggacaaagca cctccaccgg
     1321 gcctgccggt tgagaccatt aaggatctaa tgcgtcagtt tctaagcggc ctggattttc
     1381 ttcatgcaaa ctgcattgtt caccgggacc tgaagccaga gaacattcta gtgacaagta
     1441 atgggaccgt caagctggct gactttggcc tagctagaat ctacagctac cagatggccc
     1501 tcacgcctgt ggtggttacg ctctggtacc gagctcctga agttcttctg cagtctacat
     1561 acgcaacacc cgtggacatg tggagcgttg gctgtatctt tgcagagatg ttccgtcgga
     1621 agcctctctt ctgtggaaac tctgaagccg accagttggg gaaaatcttt gatctcattg
     1681 gattgcctcc agaagacgac tggcctcgag aggtatctct acctcgagga gcctttgccc
     1741 ccagagggcc tcggccagtg cagtcagtgg tgccagagat ggaggagtct ggagcgcagc
     1801 tgctactgga aatgctgacc tttaacccac ataagcgaat ctctgccttc cgagccctgc
     1861 agcactccta cctgcacaag gaggaaagcg acgcagagtg agaagagggg ctgcctttcc
     1921 cagtcttggt ggagaaaccc tcgctgaagc ggcagcctct gtttcccccc aaggctgtgg
     1981 agaatcctcc agttttttac agagaatatt ttaagcctta aataacaagt ccccacctct
     2041 ccttacgagg ttcaccccca ttaccctccc ctagctctac actaaagggc aggtgtatct
     2101 gtcttcttcc ctccctgatt tatactggga tcttttttat acaggaaaac aagacaagac
     2161 aaaaaaaaaa aaggaattcc gaggccgagt gcagaactgc tccaaaggga cctcaaggct
     2221 ttccgaggga cactaggctg actccatcga gccagtgtag agataagctt ttgtttctgt
     2281 ttatttttta tgggacccct tattgtactc ctaatgattt tgctcttcgg accctgcatt
     2341 cttaatcgat gatccaaatg agtcttcgga cctcgcgggg gccgcttaag cggtggttag
     2401 ggtttgtctg acgcgggggg agggggaagg aacgaaacac tctcattcgg aggcggctcg
     2461 gggtttggtc ttggtggcca cgggcacgca gaagagcgcc gcgatcctct taagcacccc
     2521 cccgccctcc gtggaggcgg gggtttggtc ggcgggtggt aactggcggg ccgctgactc
     2581 gggcgggtcg cgcgccccag agtgtgacct tttcggtctg ctcgcagacc cccgggcggc
     2641 gccgccgcgg cggcgacggg ctcgctgggt cctaggctcc atggggaccg tatacgtgga
     2701 caggctctgg agcatccgca cgactgcggt gatattaccg gagaccttct gcgggacgag
     2761 ccgggtcacg cggctgacgc ggagcgtccg ttgggcgaca aacaccagga cggggcacag
     2821 gtacactatc ttgtcacccg gaggcgcgag ggactgcagg agcttcaggg agtggcgcag
     2881 ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg aagaaatata
     2941 tttgcatgtc tttagttcta tgatgacaca aaccccgccc agcgtcttgt cattggcgaa
     3001 ttaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
     3061 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aacagcgtca
     3121 acagcgtgcc gcagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3961 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc
     4141 tcgatcccct cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct
     4201 accggcagtg caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg
     4261 cccccgaact gcaggagtgg ggaggcacga tggccgcttt ggtcgacatc gattagtcca
     4321 atttgttaaa gacaggatat cagtggtcca ggctctagct ttgactcaac aatatcacca
     4381 gctgaagcct atagagtacg agccatagtt aaaataaaag attttattta gtctccagaa
     4441 aaagggggga atgaaagacc ccacccgtag gtggcaagct agcttaagta acgccacttt
     4501 gcaaggcatg gaaaaataca taactgagaa tagaaaagtt cagatcaagg tcaggaacaa
     4561 agaaacagct gaataccaaa caggatatct gtggtaagcg gttcctgccc cggctcaggg
     4621 ccaagaacag atgagacagc tgagtgatgg gccaaacagg atatctgtgg taagcagttc
     4681 ctgccccggc tcggggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
     4741 tctagtgaat catcagatgt ttccagggtg ccccaaggac ctgaaaatga ccctgtacct
     4801 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttcc gctctccgag
     4861 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcttccgata gactgcgtcg
     4921 cccgggtacc cgtattccca ataaagcctc ttgctgtttg cttttttttt tttttttttt
     4981 tcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5041 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5101 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5161 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5221 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtcc
     5281 ctcgagggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     5341 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5401 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     5461 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     5521 ttatgatccg gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5581 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5641 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     5701 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5761 ataggtcgac cggtcgaccg gtcgaccggt cgacctatgc ggtgtgaaat accgcacaga
     5821 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     5881 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5941 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6001 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6061 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6121 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6181 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6241 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6301 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6361 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6421 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     6481 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6541 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6601 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6661 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6721 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6781 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6841 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6901 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6961 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7021 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7081 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7141 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7201 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7261 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7321 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7381 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7441 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7501 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7561 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7621 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7681 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7741 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7801 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     7861 ttggtcgacc aattctcatg tttgacagct tatcatcgat aagctagctt ggctgtggaa
     7921 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     7981 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     8041 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     8101 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     8161 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     8221 aggctttttt ggaggcctag gcttttgcaa aaagctcctc gatcgagggg ctcgcatctc
     8281 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     8341 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     8401 caggtcgaga ccgggccttt gtccggcgct
//
